title: A simple high-yielding process for transient gene expression in CHO cells.
abstract: Here we describe a simplified method for transient gene expression (TGE) in suspension-adapted Chinese hamster ovary (CHO) cells using polyethylenimine (PEI) for DNA delivery. Both the transfection and production phases of the bioprocess were performed at a density of 4 × 10⁶ cells/mL at 31 °C. In addition, the amounts of both PEI and plasmid DNA were reduced up to 50% on a per cell basis compared to previously published protocols from this laboratory, resulting in higher cell viability after transfection and higher volumetric recombinant protein yields. In batch cultures of up to 14 days, reproducible recombinant antibody yields up to 300 mg/L were achieved at small scale (5 mL) and up to 250 mg/L at large scale (500 mL). The simplicity and improved yields are expected to increase the utility of CHO cells for the rapid production of recombinant proteins at larger scales by TGE.
authors.affiliation: Laboratory for Cellular Biotechnology (LBTC), École Polytechnique Fédéral de Lausanne (EPFL), CH-1015 Lausanne, Switzerland.
authors.forename: Yashas
authors.initials: Y
authors.lastname: Rajendra
authors.forename: Divor
authors.initials: D
authors.lastname: Kiseljak
authors.forename: Lucia
authors.initials: L
authors.lastname: Baldi
authors.forename: David L
authors.initials: DL
authors.lastname: Hacker
authors.forename: Florian M
authors.initials: FM
authors.lastname: Wurm
mesh_terms: D000818:Animals; D001709:Biotechnology / Q000379:methods*; D016466:CHO Cells; D002452:Cell Count; D006224:Cricetinae; D003412:Cricetulus; D004247:DNA / Q000235:genetics; D015870:Gene Expression; D010957:Plasmids / Q000235:genetics; D011094:Polyethyleneimine / Q000494:pharmacology; D011994:Recombinant Proteins / Q000096:biosynthesis; D014162:Transfection
chemical_list: D011994:Recombinant Proteins; D011094:Polyethyleneimine; D004247:DNA